WB
Western blot analysis of Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) on different lysates with Rabbit anti-Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) antibody at 1/1,000 dilution.
Lane 1: NIH/3T3 cell lysate,
Lane 2: NIH/3T3 treated with 1mM sodium orthovanadate for 30 minutes cell lysate,
Lysates/proteins at 30 µg/Lane.
Exposure time: 3 minutes 10 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/100,000 dilution was used for 1 hour at room temperature.
WB
Western blot analysis of Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) on different lysates with Rabbit anti-Phospho-PI3K p85 (Y467) + PI3K p55 (Y199) antibody at 1/1,000 dilution.
Lane 1: 293T overexpress Src cell lysate,
Lane 2: 293T overexpress Src treated with 1mM sodium orthovanadate for 30 minutes cell lysate,
Lane 3: 293T overexpress Src cell lysate, the membrane treated with λpp for 1 hour,
Lane 4: 293T overexpress Src treated with 1mM sodium orthovanadate for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour.
Lysates/proteins at 20 µg/Lane.
Exposure time: 28 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/100,000 dilution was used for 1 hour at room temperature.